JPH099965A - Transgenic animal having full-length sequence of hepatitis c viral gene - Google Patents
Transgenic animal having full-length sequence of hepatitis c viral geneInfo
- Publication number
- JPH099965A JPH099965A JP7188605A JP18860595A JPH099965A JP H099965 A JPH099965 A JP H099965A JP 7188605 A JP7188605 A JP 7188605A JP 18860595 A JP18860595 A JP 18860595A JP H099965 A JPH099965 A JP H099965A
- Authority
- JP
- Japan
- Prior art keywords
- hcv
- plasmid
- full
- length cdna
- transgenic animal
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
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Links
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Landscapes
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
Description
【０００１】[0001]
【産業上の利用分野】本発明は、Ｃ型肝炎ウイルス（Ｈ
ＣＶ）全長ｃＤＮＡ配列を含むプラスミド、そのＤＮＡ
配列を含有するＤＮＡを導入した遺伝子導入動物（トラ
ンスジェニック動物）に関するものである。The present invention relates to hepatitis C virus (H
CV) A plasmid containing the full-length cDNA sequence, its DNA
The present invention relates to a transgenic animal (transgenic animal) into which a DNA containing a sequence has been introduced.
【０００２】[0002]
【従来技術及び課題】ＨＣＶは、輸血後非Ａ非Ｂ肝炎の
主な原因物質であり［Saito, I. et al., Proc. Natl.
Acad. Sci. USA., 87,6547-6549(1990)］、このウイル
スに起因した肝炎は、慢性化しやすく、肝硬変や肝癌に
移行する確率が高い。1988年にカイロン社によって、Ｈ
ＣＶのｃＤＮＡがクローニングされ［Choo, Q.-L. et a
l., Science, 244,359-362(1989)］、現在までに全塩基
配列並びに全アミノ酸配列が明らかにされた。また、Ｈ
ＣＶは、フラビウイルスやペスティウイルスに近縁なプ
ラス鎖ＲＮＡウイルスであることが明らかにされた［Ka
to, N. et al., Proc. Natl. Acad. Sci. USA., 87,952
4-9528(1990)］。BACKGROUND OF THE INVENTION HCV is a major causative agent of post-transfusion non-A non-B hepatitis [Saito, I. et al., Proc. Natl.
Acad. Sci. USA., 87, 6547-6549 (1990)], hepatitis caused by this virus is apt to become chronic and has a high probability of becoming cirrhosis or liver cancer. In 1988, by Chiron, H
CV cDNA has been cloned [Choo, Q.-L. et a
L., Science, 244, 359-362 (1989)], the whole base sequence and all amino acid sequences have been clarified so far. Also, H
CV has been revealed to be a plus-strand RNA virus closely related to flaviviruses and pestiviruses [Ka
to, N. et al., Proc. Natl. Acad. Sci. USA., 87,952
4-9528 (1990)].
【０００３】しかしながら、ＨＣＶの宿主はヒトまたは
チンパンジーに限られており、また感受性を示す培養細
胞がないことより、ウイルスタンパク質の性状やプロセ
ッシング機構の解析などの基礎研究のみならず、疫学的
な研究もあまり進歩していない。However, the host of HCV is limited to humans or chimpanzees, and since there are no cultured cells showing sensitivity, not only basic research such as analysis of properties of viral proteins and processing mechanism but also epidemiological research. Is not making much progress.
【０００４】近年、発生工学技術の発達により胚性期に
外来遺伝子を導入し、導入遺伝子を個体レベルで発現さ
せるトランスジェニックマウス作製技術が確立されてい
る［Gordon, J.W. et al., Proc. Natl. Acad. Sci. US
A., 77, 7380-7384(1980)］。この技術を用い、いくつ
かのウイルス疾患モデルマウスが確立されており、その
中には、Ｂ型肝炎の原因物質であるＢ型肝炎ウイルス
（ＨＢＶ）のＤＮＡが導入されたマウスが作製され、Ｈ
ＢＶ疾患モデル動物として利用されている［Chisari,
F.V. et al., Science, 230, 1157-1160(1985)］。しか
しながら、ＨＣＶのＤＮＡが導入されたトランスジェニ
ックマウスについては未だ報告は無い。[0004] In recent years, with the development of developmental engineering technology, a transgenic mouse production technique has been established in which a foreign gene is introduced at the embryonic stage and the transgene is expressed at the individual level [Gordon, JW et al., Proc. Natl. . Acad. Sci. US
A., 77, 7380-7384 (1980)]. Using this technology, several mouse models of viral diseases have been established. Among them, mice into which the DNA of hepatitis B virus (HBV), which is the causative agent of hepatitis B, was introduced were prepared.
It is used as a BV disease model animal [Chisari,
FV et al., Science, 230, 1157-1160 (1985)]. However, there is no report yet regarding a transgenic mouse into which HCV DNA has been introduced.
【０００５】[0005]
【課題を解決するための手段】このような状況におい
て、鋭意研究を行った結果、体細胞及び生殖細胞に、Ｈ
ＣＶ全長ｃＤＮＡが血清アミロイドＰ成分（ＳＡＰ）プ
ロモーターの下流側に挿入されているＤＮＡ配列を含有
するＤＮＡが導入されているトランスジェニック動物を
作製することに成功した。[Means for Solving the Problems] Under such circumstances, as a result of earnest research, somatic cells and germ cells were found to have
We have succeeded in producing a transgenic animal into which a DNA containing a DNA sequence in which the CV full-length cDNA is inserted downstream of the serum amyloid P component (SAP) promoter has been introduced.
【０００６】[0006]
【発明の構成】本発明において使用されるＨＣＶ遺伝子
は、本発明者らが研究に用いているＨＣＶ全長ｃＤＮＡ
が挿入されたプラスミドpHCV7由来のものが使用され
る。好ましくは、プラスミドpHCV7中に挿入されたＨＣ
Ｖ全長ｃＤＮＡのEcoRＩ−EcoRＩ領域が使用される（図
１）。DETAILED DESCRIPTION OF THE INVENTION The HCV gene used in the present invention is the full-length HCV cDNA used by the present inventors for research.
The one derived from the plasmid pHCV7 into which is inserted is used. Preferably the HC inserted in the plasmid pHCV7
The EcoRI-EcoRI region of the V full length cDNA is used (FIG. 1).
【０００７】本発明の導入遺伝子は、上記ＨＣＶ全長ｃ
ＤＮＡがヒトＳＡＰプロモーターに依存して転写される
ように構築される。ここで、ＳＡＰプロモーターは、生
後肝臓にて発現が増加するものである。このＳＡＰプロ
モーターは、プラスミドpSG-1由来のものが使用され
る。このプラスミドpSG-1は、プラスミドpAT153、ＳＡ
Ｐプロモーター及びウサギβグロビンイントロンIIより
成り、SV40のlate及びAn領域を含有する（図１）。The transgene of the present invention is the above HCV full-length c.
It is constructed so that the DNA is transcribed depending on the human SAP promoter. Here, the expression of the SAP promoter is increased in postnatal liver. As this SAP promoter, one derived from the plasmid pSG-1 is used. This plasmid pSG-1 is a plasmid pAT153, SA
It consists of the P promoter and rabbit β globin intron II and contains the late and An regions of SV40 (Fig. 1).
【０００８】ＨＣＶ全長ｃＤＮＡのEcoRＩ−EcoRＩ領域
が上記プラスミドpSG-1のウサギβグロビンイントロンI
IのEcoRＩクローニングサイトに挿入され、プラスミドp
SGHCV7が構築される。当該プラスミドpSGHCV7を組み込
んだ大腸菌Escherichia coli111927は、FERM P-15005と
して生命工学工業技術研究所に寄託されている。The EcoRI-EcoRI region of the HCV full-length cDNA is the rabbit β-globin intron I of the above plasmid pSG-1.
It was inserted into the EcoRI cloning site of I, and plasmid p
SGHCV7 is built. Escherichia coli 111927 in which the plasmid pSGHCV7 has been incorporated has been deposited with the Institute of Biotechnology, FERM P-15005.
【０００９】本発明のトランスジェニック動物を作製す
るために使用する導入遺伝子は、上記プラスミドpSGHCV
7由来のＨＣＶ全長ｃＤＮＡとＳＡＰプロモーター遺伝
子を含有するＤＮＡが使用される。好ましくは、ＳＡＰ
プロモーター遺伝子の下流に結合されたウサギβグロビ
ンイントロンII中にＨＣＶ全長ｃＤＮＡが挿入されたＤ
ＮＡ配列を含有するＤＮＡが使用される。さらに好まし
くは、pSGHCV7をHindIIIとXhoＩで消化して得られる約1
2kbのフラグメントが使用される（図２）。The transgene used for producing the transgenic animal of the present invention is the above-mentioned plasmid pSGHCV.
The HCV full-length cDNA derived from 7 and the DNA containing the SAP promoter gene are used. Preferably SAP
D with HCV full-length cDNA inserted in rabbit β-globin intron II linked downstream of the promoter gene
DNA containing the NA sequence is used. More preferably, about 1 obtained by digesting pSGHCV7 with HindIII and XhoI
A 2 kb fragment is used (Figure 2).
【００１０】得られたフラグメントを用いて、常法［K.
Yamamura et al., J. Biochem. (Tokyo), 96, 357-363
(1984)］に従い、ＨＣＶ全長ｃＤＮＡを含有するＤＮＡ
配列が導入されたトランスジェニック動物が作製され
る。使用される動物は、好ましくは齧歯動物、さらに好
ましくはマウスである。Using the obtained fragment, the conventional method [K.
Yamamura et al., J. Biochem. (Tokyo), 96, 357-363
(1984)], DNA containing HCV full-length cDNA
A transgenic animal into which the sequence has been introduced is produced. The animal used is preferably a rodent, more preferably a mouse.
【００１１】[0011]
【発明の効果】本発明において、ＨＣＶ全長ｃＤＮＡの
マウスへの導入を試みた結果、ＨＣＶ全長ＤＮＡが検出
され、さらには、肝臓においてＨＣＶ ＲＮＡの発現が
確認された。すなわち、本発明によりＨＣＶ全長遺伝子
を有するトランスジェニックマウスが確立されたことが
明らかになった。本発明により得られたＨＣＶトランス
ジェニック動物は、ＨＣＶ遺伝子機能の解析、ＨＣＶ感
染並びに感染後肝炎の発症のメカニズムの解明、さらに
は医薬品のスクリーニング等に極めて有用であると期待
される。INDUSTRIAL APPLICABILITY In the present invention, an attempt was made to introduce an HCV full-length cDNA into a mouse. As a result, HCV full-length DNA was detected, and furthermore, HCV RNA expression was confirmed in the liver. That is, it was revealed that the present invention established a transgenic mouse having a full-length HCV gene. The HCV transgenic animals obtained by the present invention are expected to be extremely useful for the analysis of HCV gene function, elucidation of the mechanism of HCV infection and development of post-infection hepatitis, and further for drug screening.
【００１２】以下、実施例にそって本発明をさらに詳細
に説明するが、これら実施例は本発明の範囲を限定する
ものではない。Hereinafter, the present invention will be described in more detail with reference to Examples, but these Examples do not limit the scope of the present invention.
【００１３】[0013]
《実施例１．導入遺伝子の調製》ＨＣＶ遺伝子は 本発
明者らが研究に用いているＨＣＶ全長ｃＤＮＡが挿入さ
れたプラスミドpHCV7由来のものを用いた（図１）。本
発明者らは導入遺伝子としてＨＣＶ全長ｃＤＮＡがヒト
血清アミロイドＰ成分（ＳＡＰ）プロモーターに依存し
て転写されるコンストラクトを作製した。このプロモー
ターは生後、肝臓にて発現が増加するもので、プラスミ
ドpSG-1由来のものを用いた。プラスミドpSG-1は、プラ
スミドpAT153、ＳＡＰプロモーター及びウサギβ−グロ
ビンイントロンIIより成り、SV40のlate及びAn領域を含
む。このプラスミドの切断部位はHindIII-XhoIで、クロ
ーニングサイトはEcoRIである。pHCV7からEcoRＩ−EcoR
Ｉで分離したＨＣＶ全長ｃＤＮＡをプラスミドpSG-1中
のウサギβグロビンイントロンII中のEcoRＩクローニン
グサイトに挿入することにより、プラスミドpSGHCV7を
得た（図１）。得られたpSGHCV7を、HindIIIとXhoＩで
消化し、最終的に12kbのフラグメントを分離し、これを
導入遺伝子フラグメントとして受精卵への注入に用いた
（図２）。本発明で導入遺伝子として使用される全ＨＣ
Ｖ遺伝子を含有するプラスミドpSGHCV7を組み込んだ大
腸菌が、Escherichia coli 111927（生命工学工業技術
研究所：FERM P-15005）として、本出願人により寄託さ
れている。<< Embodiment 1. Preparation of transgene >> The HCV gene used was derived from the plasmid pHCV7 into which the HCV full-length cDNA used in the present inventors' research was inserted (Fig. 1). The present inventors have prepared a construct in which HCV full-length cDNA is transcribed as a transgene depending on the human serum amyloid P component (SAP) promoter. Expression of this promoter is increased in the liver after birth, and the promoter derived from the plasmid pSG-1 was used. Plasmid pSG-1 consists of plasmid pAT153, the SAP promoter and rabbit β-globin intron II and contains the late and An regions of SV40. The cleavage site of this plasmid is HindIII-XhoI and the cloning site is EcoRI. pHCV7 to EcoRI-EcoR
The HCV full-length cDNA isolated in I was inserted into the EcoRI cloning site in rabbit β-globin intron II in plasmid pSG-1 to obtain plasmid pSGHCV7 (FIG. 1). The obtained pSGHCV7 was digested with HindIII and XhoI to finally separate a 12 kb fragment, which was used as a transgene fragment for injection into fertilized eggs (FIG. 2). All HC used as transgene in the present invention
Escherichia coli into which the plasmid pSGHCV7 containing the V gene has been incorporated has been deposited by the applicant as Escherichia coli 111927 (Institute of Industrial Science and Technology: FERM P-15005).
【００１４】《実施例２．遺伝子の導入》トランスジェ
ニックマウス作製にはC57BL/6マウスを用いた。常法
［K. Yamamura et al., J. Biochem. (Tokyo), 96, 357
-363(1984)］に従い、ホルモン処理した雌マウスを用
い、交配により受精卵を得、受精卵の雄性前核内に３ng
/mlに調製した導入遺伝子フラグメントをマイクロガラ
スピペットを用いて注入した。得られた遺伝子導入卵の
うち、生き残った500個の卵を偽妊娠雌マウスの卵管に
移植した（図３）。その結果、91匹のマウスが誕生し
た。Example 2 Introduction of Gene >> C57BL / 6 mice were used for producing transgenic mice. Conventional method [K. Yamamura et al., J. Biochem. (Tokyo), 96, 357
-363 (1984)], a fertilized egg was obtained by mating using a female mouse treated with hormone, and 3 ng was placed in the male pronucleus of the fertilized egg.
The transgene fragment prepared at / ml was injected using a micro glass pipette. Of the resulting transgenic eggs, 500 surviving eggs were transplanted into the oviducts of pseudopregnant female mice (Fig. 3). As a result, 91 mice were born.
【００１５】《実施例３．導入遺伝子の抽出》遺伝子導
入されたマウスより誕生したマウスの尻尾よりＤＮＡを
抽出した。マウスの尻尾をプロテイナーゼＫにより溶解
し、フェノール／クロロホルム（１：１）で処理し、エ
タノール沈殿させ、ＴＥ（10mM Tris-HCl(pH8.0)/1mM E
DTA）中に再溶解した。Example 3 Extraction of Transgene >> DNA was extracted from the tail of a mouse born from a transgenic mouse. The tail of the mouse was dissolved with proteinase K, treated with phenol / chloroform (1: 1), ethanol-precipitated, and TE (10 mM Tris-HCl (pH8.0) / 1 mM E was added).
Redissolved in DTA).
【００１６】《実施例４．導入遺伝子の解析》導入遺伝
子の解析は、サザンブロット解析によって行った。実施
例３で得られたマウス尾ＤＮＡ抽出液（10μg）をEcoR
Ｉで消化後 0.75% アガロースゲルで電気泳動し、ナイ
ロン膜（hybond-N+）に0.1Ｍ NaCl／0.1Ｍ NaOHでアル
カリトランスファーした。ハイブリダイゼーションは、
コア領域またはNS5領域（図２）をディゴキシゲニンで
標識（DIG-DNA標識キット／ベーリンガーマンハイム
社）したプローブを用いて行い、化学発光（CSPD／ベー
リンガーマンハイム社）または発色（NTB/BCIP／ベーリ
ンガーマンハイム社）によって検出を行った。Example 4 Analysis of transgene >> Analysis of the transgene was performed by Southern blot analysis. The mouse tail DNA extract (10 μg) obtained in Example 3 was EcoR
After digestion with I, electrophoresis was carried out on 0.75% agarose gel, and alkaline transfer was performed on a nylon membrane (hybond-N +) with 0.1M NaCl / 0.1M NaOH. Hybridization is
Chemiluminescence (CSPD / Boehringer Mannheim) or color development (NTB / BCIP / Boehringer Mannheim) was performed using a probe in which the core region or NS5 region (Fig. 2) was labeled with digoxigenin (DIG-DNA labeling kit / Boehringer Mannheim). ).
【００１７】その結果、16匹のマウスにおいて、図４に
示したように、9.5kbp付近に単一バンドが認められた。
この結果から、これら16匹のマウスは完全な長さのＨＣ
Ｖ遺伝子を保有していることが確認された。これらのマ
ウスをＦ0（創生）マウスと呼ぶ。得られた各々のトラ
ンスジェニックマウスについて導入遺伝子のコピー数の
解析を行った結果、１コピーから100コピー以上の導入
遺伝子をもつマウスの系統が得られた（表１）。As a result, in 16 mice, as shown in FIG. 4, a single band was recognized around 9.5 kbp.
From this result, these 16 mice showed full length HC
It was confirmed to carry the V gene. These mice are called F0 (creative) mice. As a result of analyzing the number of copies of the transgene in each of the obtained transgenic mice, a mouse strain having 1 to 100 or more copies of the transgene was obtained (Table 1).
【００１８】[0018]
【表１】 ※死亡：いずれも交配中の死亡[Table 1] * Death: Death during mating
【００１９】《実施例５．ＨＣＶトランスジェニックマ
ウスの系統確立》得られたＨＣＶトランスジェニックマ
ウス（Ｆ0）の子孫を得るために同系統であるC57BL/6の
Wild typeと交配を行った。その結果、10系統が確立さ
れた（表１）。Example 5 Establishment of strain of HCV transgenic mouse >> To obtain progeny of the obtained HCV transgenic mouse (F0)
I crossed with Wild type. As a result, 10 strains were established (Table 1).
【００２０】《実施例６．ＨＣＶトランスジェニックマ
ウスのＲＮＡ発現解析》実施例５において系統化された
系統４及び70について、子孫１代目（Ｆ1）または２代
目（Ｆ2）のマウスを用いてＲＮＡの発現の有無を確認
した。解析は、ＲＰＡIIキット（アンビオン社製）を用
いて、添付のプロトコールに従って、リボヌクレアーゼ
・プロテクション・アッセイを行った。サンプルは、マ
ウスの肝臓を摘出し、ＩＳＯＧＥＮ（ニッポンジーン
社）を用いてトータルＲＮＡを得た。プローブは、ＨＣ
ＶｃＤＮＡをNcoＩ−PvuIIで消化した1429から1714ntの
領域を鋳型にし、32Ｐで標識したＲＮＡプローブ（PENS
1-300）をin vitro転写によって作製した。プローブは
結果として、in vitro転写に用いたプラスミド由来の45
bpを持つ。この領域がＨＣＶＲＮＡと非ハイブリダイズ
領域となる。サンプルＲＮＡとプローブを２日間ハイブ
リダイズを行った後、リボヌクレアーゼＡ及びＴ１によ
って消化し、５％アクリルアミド−８Ｍ尿素ゲルで電気
泳動し、乾燥後、オートラジオグラフィーを行った。そ
の結果、今回解析を実施した２系統、系統４及び70にお
いて肝臓でのＨＣＶＲＮＡの発現が確認された（図
５）。Example 6. RNA Expression Analysis of HCV Transgenic Mice >> For the lines 4 and 70 systematized in Example 5, the presence or absence of RNA expression was confirmed using the progeny first generation (F1) or second generation (F2) mice. For the analysis, a ribonuclease protection assay was performed using an RPAII kit (manufactured by Ambion) according to the attached protocol. As a sample, mouse liver was extracted and total RNA was obtained using ISOGEN (Nippon Gene). The probe is HC
An RNA probe (PENS) labeled with 32 P using the 1429 to 1714 nt region obtained by digesting VcDNA with NcoI-PvuII as a template.
1-300) was prepared by in vitro transcription. The probe resulted in 45 plasmids derived from the plasmid used for in vitro transcription.
has bp. This region becomes a non-hybridizing region with HCV RNA. After the sample RNA and the probe were hybridized for 2 days, they were digested with ribonuclease A and T1, electrophoresed on a 5% acrylamide-8M urea gel, dried, and then autoradiographed. As a result, expression of HCV RNA in the liver was confirmed in the two strains analyzed this time, strains 4 and 70 (Fig. 5).
【図１】 ＨＣＶ導入遺伝子の構築図を示す。FIG. 1 shows a construction diagram of an HCV transgene.
【図２】 トランスジェニック動物作製に使用されるＨ
ＣＶ導入遺伝子及びプローブ領域を示す。FIG. 2 H used for producing transgenic animals
The CV transgene and probe region are shown.
【図３】 トランスジェニックマウスの作製方法を示
す。FIG. 3 shows a method for producing a transgenic mouse.
【図４】 サザンブロット分析によりＦ0マウスのＨＣ
Ｖ遺伝子導入の確認した結果を示す。FIG. 4: HC of F0 mice by Southern blot analysis
The confirmed result of V gene introduction is shown.
【図５】 リボヌクレアーゼプロテクション分析によ
り、ＨＣＶトランスジェニックマウスの肝臓でのＲＮＡ
の発現を確認した結果を示す。FIG. 5. RNA in liver of HCV transgenic mice by ribonuclease protection analysis.
The result of having confirmed the expression of is shown.
Claims (8)
Ａが血清アミロイドＰ成分（ＳＡＰ）プロモーター遺伝
子の下流側に挿入されているＤＮＡ配列を含有すること
を特徴とするプラスミド。1. A hepatitis C virus (HCV) full-length cDNA
A plasmid characterized in that A contains a DNA sequence inserted downstream of the serum amyloid P component (SAP) promoter gene.
ーター遺伝子の下流側に結合されたウサギβグロビンイ
ントロンII領域中に挿入されているＤＮＡ配列を含有す
ることを特徴とする請求項１記載のプラスミド。2. The plasmid according to claim 1, wherein the HCV full-length cDNA contains a DNA sequence inserted in the rabbit β-globin intron II region linked to the downstream side of the SAP promoter gene.
て、寄託番号FERM P-15005として寄託されている請求項
１または２のいずれかに記載のプラスミド。3. The plasmid according to claim 1 or 2, which has been incorporated into Escherichia coli and deposited under the deposit number FERM P-15005.
ＮＡがＳＡＰプロモターの下流側に挿入されているＤＮ
Ａ配列を含むＤＮＡが導入されているトランスジェニッ
ク動物。4. HCV full-length cDNA in somatic cells and germ cells
DN in which NA is inserted downstream of the SAP promoter
A transgenic animal into which a DNA containing an A sequence has been introduced.
ーター遺伝子の下流側に結合されたウサギβグロビンイ
ントロンII領域中に挿入されているＤＮＡ配列を含むＤ
ＮＡが導入されていることを特徴とする請求項４記載の
トランスジェニック動物。5. A D containing the DNA sequence in which the HCV full-length cDNA is inserted into a rabbit β-globin intron II region linked downstream of the SAP promoter gene.
The transgenic animal according to claim 4, wherein NA is introduced.
記載のプラスミドのHindIII−XhoＩ領域であることを特
徴とする請求項４または５に記載のトランスジェニック
動物。6. A DNA containing the DNA sequence according to claim 1.
The transgenic animal according to claim 4 or 5, which is the HindIII-XhoI region of the described plasmid.
する請求項４から６のいずれかに記載のトランスジェニ
ック動物。7. The transgenic animal according to any one of claims 4 to 6, wherein the animal is a rodent.
とする請求項４から７のいずれかに記載のトランスジェ
ニック動物。8. The transgenic animal according to claim 4, wherein the rodent is a mouse.
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|---|---|---|---|
| JP18860595A JP3714702B2 (en) | 1995-06-30 | 1995-06-30 | Transgenic animal having full-length sequence of hepatitis C virus gene |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP18860595A JP3714702B2 (en) | 1995-06-30 | 1995-06-30 | Transgenic animal having full-length sequence of hepatitis C virus gene |
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| Publication Number | Publication Date |
|---|---|
| JPH099965A true JPH099965A (en) | 1997-01-14 |
| JP3714702B2 JP3714702B2 (en) | 2005-11-09 |
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ID=16226603
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP18860595A Expired - Fee Related JP3714702B2 (en) | 1995-06-30 | 1995-06-30 | Transgenic animal having full-length sequence of hepatitis C virus gene |
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| Country | Link |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1999067394A1 (en) | 1998-06-24 | 1999-12-29 | Chugai Seiyaku Kabushiki Kaisha | Vector expressing the full-length gene of rna virus and use thereof |
| WO2003037081A1 (en) * | 2001-10-30 | 2003-05-08 | Tokyo Metropolitan Organization For Medical Research | Hcv gene transgenic animal |
| EP1362510A3 (en) * | 2002-05-17 | 2003-11-26 | Beijing Institute of Biotechnology | A mouse model for hepatocellular carcinoma by targeted integration of hepatitis B virus genes |
| JP2007530072A (en) * | 2004-04-01 | 2007-11-01 | ニューロテック ファーマシューティカルズ カンパニー リミテッド | Alzheimer's disease-induced transformed mouse expressing mutant βCTF99 |
| JP2008537042A (en) * | 2005-04-19 | 2008-09-11 | ベーエスハー ボッシュ ウント ジーメンス ハウスゲレーテ ゲゼルシャフト ミット ベシュレンクテル ハフツング | Household equipment having a locking device |
-
1995
- 1995-06-30 JP JP18860595A patent/JP3714702B2/en not_active Expired - Fee Related
Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1999067394A1 (en) | 1998-06-24 | 1999-12-29 | Chugai Seiyaku Kabushiki Kaisha | Vector expressing the full-length gene of rna virus and use thereof |
| WO2003037081A1 (en) * | 2001-10-30 | 2003-05-08 | Tokyo Metropolitan Organization For Medical Research | Hcv gene transgenic animal |
| JPWO2003037081A1 (en) * | 2001-10-30 | 2005-02-17 | 財団法人 東京都医学研究機構 | HCV gene transgenic animals |
| EP1362510A3 (en) * | 2002-05-17 | 2003-11-26 | Beijing Institute of Biotechnology | A mouse model for hepatocellular carcinoma by targeted integration of hepatitis B virus genes |
| JP2010200756A (en) * | 2002-05-17 | 2010-09-16 | Beijing Inst Of Biotechnology | Mouse model for inducing hepatocellular carcinoma by incorporation targeting target of hepatitis b virus gene |
| JP2007530072A (en) * | 2004-04-01 | 2007-11-01 | ニューロテック ファーマシューティカルズ カンパニー リミテッド | Alzheimer's disease-induced transformed mouse expressing mutant βCTF99 |
| JP2008537042A (en) * | 2005-04-19 | 2008-09-11 | ベーエスハー ボッシュ ウント ジーメンス ハウスゲレーテ ゲゼルシャフト ミット ベシュレンクテル ハフツング | Household equipment having a locking device |
| US8377230B2 (en) | 2005-04-19 | 2013-02-19 | Bsh Bosch Und Siemens Hausgeraete Gmbh | Household appliance comprising a locking device |
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| Publication number | Publication date |
|---|---|
| JP3714702B2 (en) | 2005-11-09 |
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